摘要
从患病华南虎分离病原菌,进行快速鉴定,同时以大肠杆菌16S rRNA基因的通用引物和irp2、papC、iucD、tsh、iss毒力基因的特异性引物进行PCR扩增、测序,并将扩增出来的irp2、iucD、iss基因序列与Genbank相应序列进行同源性分析。测序鉴定为大肠埃希氏菌,与传统细菌鉴定方法结果相一致,分离菌携带irp2、iucD、iss毒力基因,从毒力试验得到证实。其序列与Genbank上发表的iucD、iss基因序列同源性分别高达98%、99%,而irp2基因序列的同源性仅为48%。采用16S rRNA基因序列分析法可以对华南虎大肠埃希氏菌感染进行快速鉴定,华南虎源性大肠杆菌同时携带有irp2、iucD、iss 3个毒力基因,可能与其致病性有关系。
Our research was aimed at rapid identification of pathogenic bacteria isolated from South China Tiger(Panthera tigris amoyensis).A universal primer - pair of the 16S rRNA gene and specific primer - pairs irp2,papC,iucD,tsh,and iss virulence genes were used to amplify target DNA fragments.Amplified products were sequenced and compared for homology with their sequences in Genbank.According to the sequence obtained,the isolated pathogen was identified as E.coli,which was the same as the result from biochemical tests.The isolated pathogen carried irp2,iucD,and iss genes,as confirmed by the toxicity test.Sequence homology between the iucD gene of the isolated pathogen and relevant sequences of iucD in Genbank was 98% and that of the iss gene was 99%,while that of the irp2 gene was 48%.16S rRNA gene sequence analysis can be used to rapidly detect Escherichia coli of South China Tiger,and the irp2,iucD,and iss virulence genes carried by Escherichia coli of South China Tiger may contribute to pathogenicity.
出处
《野生动物》
2012年第3期109-112,133,共5页